PT  - JOURNAL ARTICLE
AU  - William J. Nicolas
AU  - Debnath Ghosal
AU  - Elitza I. Tocheva
AU  - Elliot M. Meyerowitz
AU  - Grant J. Jensen
TI  - Structure of the bacterial cellulose ribbon and its assembly-guiding cytoskeleton by electron cryotomography
AID  - 10.1101/2020.04.16.045534
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.04.16.045534
4099  - http://biorxiv.org/content/early/2020/04/17/2020.04.16.045534.short
4100  - http://biorxiv.org/content/early/2020/04/17/2020.04.16.045534.full
AB  - Cellulose is a widespread component of bacterial biofilms, where its properties of exceptional water retention, high tensile strength and stiffness prevents dehydration and mechanical disruption of the biofilm. Bacteria in the Gluconacetobacter genus secrete crystalline cellulose, with a structure very similar to that found in plant cell walls. How this higher-order structure is produced is poorly understood. We used cryo-electron tomography and focused ion beam milling of native bacterial biofilms to image cellulose-synthesizing G. hansenii and G. xylinus bacteria in a frozen-hydrated, near-native state. We confirm previous results suggesting that cellulose crystallization occurs serially following its secretion along one side of the cell, leading to a cellulose ribbon that can reach several microns in length and combine with ribbons from other cells to form a robust biofilm matrix. We were able to take direct measurements in a near-native state of the cellulose sheets. Our results also reveal a novel cytoskeletal structure, that we name the cortical belt, adjacent to the inner membrane and underlying the sites where cellulose is seen emerging from the cell. We find that this structure is not present in other cellulose-synthesizing bacterial species, Agrobacterium tumefaciens and Escherichia coli 1094, which do not produce organized cellulose ribbons. We therefore propose that the cortical belt holds the cellulose synthase complexes in a line, to form higher-order cellulose structures such as sheets and ribbons.Competing Interest StatementThe authors have declared no competing interest.